Viral vectored vaccines for swine influenza virus and methods of use

Abstract

Provided herein are genetically engineered Pichinde viruses that include three ambisense genomic segments. Two of the genomic segments include an additional coding region that may encode one or more swine influenza virus (SIV) proteins, such as a SIV hemagglutinin (HA) protein. SIV HA proteins can be from different subtypes of SIV. Also provided herein is a reverse genetics system for making genetically engineered Pichinde virus, and a collection of vectors that can be used to produce genetically engineered Pichinde virus. Further provided are methods for using a reverse genetics system, and methods for treating a SIV infection in a subject.

1 . A genetically engineered Pichinde virus comprising:
 three ambisense genomic segments,
 wherein the first genomic segment comprises a coding region encoding a Z protein and a coding region encoding an L RdRp protein, 
 wherein the second genomic segment comprises a coding region encoding a nucleoprotein (NP) and a second coding region, wherein the second coding region encodes at least one SIV protein, and 
 wherein the third genomic segment comprises a coding region encoding a glycoprotein and a third coding region, wherein the third coding region encodes at least one SIV protein. 
   
     
     
         2 . The virus of  claim 1  wherein the at least one SIV protein encoded by the second coding region is different than the at least one SIV protein encoded by the third coding region. 
     
     
         3 . The virus of  claim 1  wherein the at least one SIV protein encoded by the second coding region is the same as the at least one SIV protein encoded by the third coding region. 
     
     
         4 . The virus of  claim 1  wherein the second coding region, the third coding region, or both encode a polycistronic message encoding at least two SIV proteins. 
     
     
         5 . The virus of  claim 4  wherein the SIV proteins encoded by the second coding region are different than the SIV proteins encoded by the third coding region. 
     
     
         6 . The virus of  claim 4  wherein the SIV proteins encoded by the second coding region are the same as the SIV proteins encoded by the third coding region. 
     
     
         7 . The virus of  claim 1  wherein the second coding region, the third coding region, or both express a monocistronic message encoding at least two SIV proteins, wherein the monocistronic message comprises nucleotides encoding a self-cleaving peptide, and wherein the nucleotides are located between the SIV proteins. 
     
     
         8 . The virus of  claim 7  wherein the self-cleaving peptide comprises a 2A peptide. 
     
     
         9 . The virus of  claim 1  wherein the at least one SIV protein of the second genomic segment and the at least one SIV protein of the third genomic segment are selected from SIV hemagglutinin (HA) proteins. 
     
     
         10 . The virus of  claim 9  wherein the HA proteins are selected from a HA expressed by a SIV subtype H1N1, a HA expressed by a SIV subtype H1N2, or a HA expressed by a SIV subtype H3N2. 
     
     
         11 . An infectious virus particle comprising the three genomic segments of  claim 1 . 
     
     
         12 . A composition comprising an isolated infectious virus particle of  claim 11 . 
     
     
         13 . A collection of vectors comprising:
 a first vector encoding the first genomic segment of  claim 1 , wherein the first genomic segment is antigenomic,   a second vector encoding the second genomic segment of  claim 1 , wherein the second genomic segment is antigenomic, and   a third vector encoding the third genomic segment of  claim 1 , wherein the third genomic segment is antigenomic.   
     
     
         14 . A method for making a genetically engineered Pichinde virus comprising:
 introducing into a cell the collection of vectors of claim  13 ; and   incubating the cells in a medium under conditions suitable for expression and packaging of the first, second, and third genomic segments.   
     
     
         15 . A reverse genetics system for making a genetically engineered virus comprising three vectors,
 wherein a first vector encodes the first genomic segment of  claim 1 , wherein the first genomic segment is antigenomic,   wherein the second vector encodes the second genomic segment of  claim 1 , wherein the second genomic segment is antigenomic, and   wherein the third vector encodes the third genomic segment of  claim 1 , wherein the third genomic segment is antigenomic.   
     
     
         16 . A method for using a reverse genetics system, comprising:
 introducing into a cell the three vectors of genomic segments of claim  15 ; and   incubating the cell under conditions suitable for transcription of the three genomic segments and expression of the coding regions of each genomic segment.   
     
     
         17 . A method for producing an immune response in a subject, comprising:
 administering to a subject the infectious virus particle of  claim 11 .   
     
     
         18 . The method of  claim 17  wherein the subject is a porcine animal. 
     
     
         19 . The method of  claim 17  wherein the immune response comprises a humoral immune response. 
     
     
         20 . The method of  claim 17  wherein the immune response comprises a cell-mediated immune response.